LOCUS pTM 6862 bp DNA circular SYN 27-OCT-2020 DEFINITION Synthetic plasmid. ACCESSION pTM VERSION KEYWORDS . SOURCE Synthetic plasmid ORGANISM Synthetic plasmid other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 6862) AUTHORS Sinegubova,M.V., Orlova,N.A. and Vorobiev,I.I. TITLE Efficient production of SARS-CoV-2 S-protein receptor binding domain (RBD) in the stably transfected CHO cells JOURNAL unpublished REFERENCE 2 (bases 1 to 6862) AUTHORS Sinegubova,M.V., Orlova,N.A. and Vorobiev,I.I. TITLE Direct Submission JOURNAL Submitted (27-OCT-2020) lab of Mammalian Cells Bioengineering, Institute of Bioengineering of The Federal Research Centre 'Fundamentals of Biotechnology' of the Russian Academy of Sciences, 60 let Oktjabrja pr-t, 7, bld. 1, Moscow, 117312, Russia COMMENT Bankit Comment: Vecscreen Comment:Submitter says that this sequence is a Synthetic Construct Bankit Comment: ALT EMAIL:ptichman@gmail.com Bankit Comment: TOTAL # OF SEQS:1. FEATURES Location/Qualifiers source 1..6862 /organism="Synthetic plasmid" /mol_type="other DNA" /db_xref="taxon:1440148" CDS 15..>83 /codon_start=1 /transl_table=11 /product="human TPA leader peptide" /translation="MDAMKRGLCCVLLLCGAVFVSAS" CDS <93..>122 /codon_start=1 /transl_table=11 /product="c-myc tag" /translation="EQKLISEEDL" CDS <135..167 /codon_start=1 /transl_table=11 /product="10xHis tag" /translation="HHHHHHHHHH" regulatory 184..771 /regulatory_class="ribosome_binding_site" /note="internal ribosomal entry site of EMCV" CDS 784..1347 /EC_number="1.5.1.3" /note="dihydrofolate reductase" /codon_start=1 /transl_table=11 /product="DHFR" /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSV EGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIE QPELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLL PEYPGVLSEVQEEKGIKYKFEVYEKKD" regulatory 1349..1963 /regulatory_class="other" /note="CHO EEF1A DFR; downstream flanking area of the EEF1A1 gene" regulatory 1614..1620 /regulatory_class="polyA_signal_sequence" regulatory complement(2128..2801) /regulatory_class="replication_regulatory_region" /note="pUC replication origin" CDS complement(2943..3803) /EC_number="3.5.2.6" /note="ampicillin resistance protein" /codon_start=1 /transl_table=11 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" regulatory complement(3798..3902) /regulatory_class="promoter" /note="bla promoter" regulatory complement(3938..4339) /regulatory_class="other" /note="EBVTR, fragment of the long terminal repeat from the Epstein-Barr virus" regulatory 4349..6860 /regulatory_class="other" /note="CHO EEF1A UFR; Upstream flanking area of the EEF1A1 gene" regulatory 5842..5847 /regulatory_class="TATA_box" BASE COUNT 1618 a 1806 c 1878 g 1560 t ORIGIN 1 tcgaggccgc caccatggat gcaatgaaga gagggctctg ctgtgtgctg ctgctgtgtg 61 gagcagtctt cgtctcggct agcgctaccg gtgaacaaaa actgatcagc gaagaggatc 121 tgtctgcagg cggtcatcac catcaccatc accatcacca tcactagcgg ccgcgcagtt 181 aacgccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag 241 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga 301 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg 361 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt 421 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca 481 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc 541 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat 601 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc 661 ctcggtacac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga 721 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aagatctgcc 781 accatggttc gaccattgaa ctgcatcgtc gccgtgtccc aaaatatggg gattggcaag 841 aacggagacc taccctggcc tccgctcagg aacgagttca agtacttcca aagaatgacc 901 acaacctctt cagtggaagg taaacagaat ctggtgatta tgggtaggaa aacctggttc 961 tccattcctg agaagaatcg acctctaaag gacagaatta atatagttct cagtagagaa 1021 ctcaaagaac caccacgagg agctcatttt cttgccaaaa gtttggatga tgccttaaga 1081 cttattgaac aaccggaatt ggcaagtaaa gtagacatgg tttggatagt cggaggcagt 1141 tctgtttacc aggaagccat gaatcaacca ggccacctca gactctttgt gacaaggatc 1201 atgcaggaat ttgaaagtga cacgtttttc ccagaaattg atttggggaa atataaactt 1261 ctcccagaat acccaggcgt cctctctgag gtccaggagg aaaaaggcat caagtataag 1321 tttgaagtct acgagaagaa agactaatca tattacccct aacacctgcc accccagtct 1381 taatcagtgg tggaagaacg gtctcagaac tgtttgtctc aattggccat ttaagtttaa 1441 tagtgaaaga ctggttaatg ataacaatgc atcggaaaac cttcaggagg aaaggagaat 1501 gttttgtgga acatttttgt gtgtgtggca gttttaagtt attagttttc aaaatcagta 1561 ctttttaatg gaaacaactt gaccaaaaat ctgtcacaga attttgagac ccattaaaat 1621 acaagtttaa tgagaagtct gtctctgtta atgctgaagt cattactaag tgcttagctt 1681 agcaaggtat gtggatgccc atttgtgttc caagggattg gactgttcat caggacccag 1741 agctgagttt caagggctca agagatggct tattacctgt gggtgtcttg aaggttctgg 1801 ttgggacaaa ttaggaatgt ttttggcaga catggtgact acccttcatc tgggtgagtt 1861 cagttgattt gtcttgagcc tttggggttt acacaagtaa atgacatcat acagttagtg 1921 tattgttagt gaatattaat atatgaggca ggctttgctc tagctagaca tatgtcgacg 1981 tccggagctc aggcgcgcca tggctcttcc gcttcctcgc tcactgactc gctgcgctcg 2041 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 2101 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 2161 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 2221 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 2281 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 2341 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat 2401 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 2461 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 2521 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 2581 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt 2641 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 2701 aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa 2761 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 2821 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 2881 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 2941 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 3001 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 3061 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 3121 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 3181 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 3241 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 3301 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 3361 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 3421 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 3481 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 3541 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 3601 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 3661 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 3721 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 3781 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 3841 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 3901 gttccgcgca catttccccg aaaagtgcca cctgcatctc atcacggtca cgcatggctg 3961 cgcccgcgca gcgccccccg accccccccc gatccccgga acgccagccg ccatccaccg 4021 cccgcagccc ccggagcccg cggaccccga ccccccgccg ctcgcccgcc gttgcgccgc 4081 tctctgcggg ggggctaggg ggccgcgggg ggaaggccac gccccctcca ctttttccag 4141 gaatgcgcgg catgacccgc cccccccatg acccgccaac acccccccga tgccccccgg 4201 gggtctttcc tggggggcct ttgtcagggt tgcctgtgtc acccacgggc cccttggcca 4261 cgcttgcctg ggtcaccccg gagccccggg tcacgtttgc cggggtcagc cctggggcgc 4321 ttcttcacgc tccattgcca tgcagaattc ccacatgtgg acaccggaca gtaggtcctc 4381 aaatgctcct tattaggttg gttcaataat atcaattgtt tgttactagg cagtgatgtt 4441 gtacatctgg aggagatctc ttgagcccat aatcaggtta ttaggaataa atactctaag 4501 gctaaaaatg tagcttagtg ataagagtgc ttgcctggtg tgctgagacc ctcggttcca 4561 tctccacaac cccatattcc attacaaaat accttttcac cgtccctagc attaagaaac 4621 aaaacaacaa agaagttttt ctttcttctg agatcctgcc cggagaggca tttaaaactg 4681 gccagggcca aaaaaaaaaa aaaaaaaaga aaaaaaagaa aagaaaacag gctagggccg 4741 gcatggtggc gcacgccttt aatcccagca cgcaggaggc agaggcaggg cggatctctg 4801 tgagtttgag gtcagcctgg tctacctagt gagtttcagg gcacccaggg ctaaagagac 4861 tgtctcaaaa acaaaacagc cacacaatca gaaccacagc aaaacgcagt tatgatcctt 4921 ggaactgtag gaatgacaag catttaaata ataggacgag ccatttttga gaagctctga 4981 tttcacaagt gtcagggatg ggctctgggc gagtaagatt gctaatgctg gcctctaaat 5041 gagaccacgt ggagttgatt agattctttt catgttcctc gtgctctatc aaataactgt 5101 acccaaatac acacacacac acacacacac aatgcgcgca cacacaaaat ccttttttag 5161 cttaagaagc ccagaatcag aagtaaagct aactgtggga cttaagtatt attctgaacg 5221 gaactcccag ggcgtgaagc gcgcttcagg cttccagaga agcagctggc gctggatgga 5281 atgaaccaag aggccagcac aggggcagat ccgtcgagct ctcggccacc gagctgagcc 5341 cttaggttct ggggctggga agggtcccta ggattgtgca cctctcccgc gggggacaag 5401 cagggggatg gcggggctga cgtcgggagg tggcctccac gggaagggac acccggatct 5461 cgacacagcc ttggcagtgg agtcaggaag ggtagggaca gattctggac gccctcttgg 5521 ccagctcctc accgccccac ccccgactgg agccgagagt aattcataca aaaggaggga 5581 tcgccttcgc ccctgggaat cccagggacc gtcgctaaat tctggccggc ctcccagccc 5641 ggaaccgctg tgcccgccca gcgcggcggg aggagcctgc gcctagggcg gatcgcgggt 5701 cggcgggaga gcacaagccc acagtccccg gcggtggggg aggggcgcgc tgagcggggg 5761 cccgggagcc agcgcggggc aaactgggaa agtggtgtcg tgtgctggct ccgccctctt 5821 cccgagggtg ggggagaacg gtataaaagt gcggtagtcg cgttggacgt tctttttcgc 5881 aacgggtttg ccgtcagaac gcaggtgagt ggcgggtgtg gcctccgcgg gcccgggctc 5941 cctcctttga gcggggtcgg accgccgtgc gggtgtcgtc ggccgggctt ctctgcgagc 6001 gttcccgccc tggatggcgg gctgtgcggg agggcgaggg ggggaggcct ggcggcggcc 6061 ccggagcctc gcctcgtgtc gggcgtgagg cctagcgtgg cttccgcccc gccgcgtgcc 6121 accgcggccg cgctttgctg tctgcccggc tgccctcgat tgcctgcccg cggcccgggc 6181 caacaaaggg agggcgtgga gctggctggt agggagcccc gtagtccgca tgtcgggcag 6241 ggagagcggc agcagtcggg ggggggaccg ggcccgcccg tcccgcagca catgtccgac 6301 gccgcctgga cgggtagcgg cctgtgtcct gataaggcgg ccgggcggtg ggttttagat 6361 gccgggttca ggtggccccg ggtcccggcc cggtctggcc agtaccccgt agtggcttag 6421 ctccgaggag ggcgaggccc gcccgcccgg caccagttgc gtgcgcggaa agatggccgc 6481 tcccgggccc tgtagcaagg agctcaaaat ggaggacgcg gcagcccggc ggagcggggc 6541 gggtgagtca cccacacaaa ggaagagggc cttgcccctc gccggccgct gcttcctgtg 6601 accccgtggt gtaccggccg cacttcagtc accccgggcg ctctttcgga gcaccgctgg 6661 cctccgctgg gggaggggat ctgtctaatg gcgttggagt ttgctcacat ttggtgggtg 6721 gagactgtag ccaggccagc ctggccatga aagtaattct tggaatttgc ccattttgag 6781 tttggagcga agctgattga caaagctgct tagccgttca aaggtattct tcgaactttt 6841 tttttaaggt gttgtgaaaa cc //